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Assessing the Suppressive Activity of Foxp3+ Regulatory T Cells

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T-Helper Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1193))

Abstract

Foxp3+ regulatory T cells (Tregs) balance the mammalian immune system by mechanisms that are yet to be elucidated in their entirety. Methods employed to quantify the regulatory activity of Tregs in vitro are an important tool in cellular immunology, but can be technically demanding and subjected to variation. In this manuscript, we describe in detail a robust Treg suppression assay based on the flow cytometric quantification of both CD4+ and CD8+ effector T cell functions. This method can provide valuable insights into the immunosuppressive activity of Foxp3+ Tregs and is versatile with regard to genetic or pharmacologic manipulations. Additionally, novel regulatory immune cells can be characterized by using this assay.

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References

  1. Mayer CT, Berod L, Sparwasser T (2012) Layers of dendritic cell-mediated T cell tolerance, their regulation and the prevention of autoimmunity. Front Immunol 3:183. doi:10.3389/fimmu.2012.00183

    PubMed Central  PubMed  Google Scholar 

  2. Berod L, Puttur F, Huehn J, Sparwasser T (2012) Tregs in infection and vaccinology: heroes or traitors? Microb Biotechnol 5(2):260–269. doi:10.1111/j.1751-7915.2011.00299.x

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Klages K, Mayer CT, Lahl K, Loddenkemper C, Teng MW, Ngiow SF, Smyth MJ, Hamann A, Huehn J, Sparwasser T (2010) Selective depletion of Foxp3+ regulatory T cells improves effective therapeutic vaccination against established melanoma. Cancer Res 70(20):7788–7799. doi:10.1158/0008-5472.CAN-10-1736, 0008-5472.CAN-10-1736 [pii]

    Article  CAS  PubMed  Google Scholar 

  4. Takahashi T, Kuniyasu Y, Toda M, Sakaguchi N, Itoh M, Iwata M, Shimizu J, Sakaguchi S (1998) Immunologic self-tolerance maintained by CD25 + CD4+ naturally anergic and suppressive T cells: induction of autoimmune disease by breaking their anergic/suppressive state. Int Immunol 10(12):1969–1980

    Article  CAS  PubMed  Google Scholar 

  5. Thornton AM, Shevach EM (1998) CD4 + CD25+ immunoregulatory T cells suppress polyclonal T cell activation in vitro by inhibiting interleukin 2 production. J Exp Med 188(2):287–296

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Lahl K, Loddenkemper C, Drouin C, Freyer J, Arnason J, Eberl G, Hamann A, Wagner H, Huehn J, Sparwasser T (2007) Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. J Exp Med 204(1):57–63. doi:10.1084/jem.20061852, jem.20061852 [pii]

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Kim J, Lahl K, Hori S, Loddenkemper C, Chaudhry A, deRoos P, Rudensky A, Sparwasser T (2009) Cutting edge: depletion of Foxp3+ cells leads to induction of autoimmunity by specific ablation of regulatory T cells in genetically targeted mice. J Immunol 183(12):7631–7634. doi:10.4049/jimmunol.0804308, jimmunol.0804308 [pii]

    Article  CAS  PubMed  Google Scholar 

  8. Mayer CT, Kuhl AA, Loddenkemper C, Sparwasser T (2012) Lack of Foxp3+ macrophages in both untreated and B16 melanoma-bearing mice. Blood 119(5):1314–1315. doi:10.1182/blood-2011-11-392266, 119/5/1314 [pii]

    Article  CAS  PubMed  Google Scholar 

  9. Mayer CT, Floess S, Baru AM, Lahl K, Huehn J, Sparwasser T (2011) CD8+ Foxp3+ T cells share developmental and phenotypic features with classical CD4+ Foxp3+ regulatory T cells but lack potent suppressive activity. Eur J Immunol 41(3):716–725. doi:10.1002/eji.201040913

    Article  CAS  PubMed  Google Scholar 

  10. Baru AM, Untucht C, Ganesh V, Hesse C, Mayer CT, Sparwasser T (2012) Optimal isolation of functional Foxp3+ induced regulatory T cells using DEREG mice. PLoS One 7(9):e44760. doi:10.1371/journal.pone.0044760

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Zal T, Volkmann A, Stockinger B (1994) Mechanisms of tolerance induction in major histocompatibility complex class II-restricted T cells specific for a blood-borne self-antigen. J Exp Med 180(6):2089–2099

    Article  CAS  PubMed  Google Scholar 

  12. Lutz MB, Kukutsch N, Ogilvie AL, Rossner S, Koch F, Romani N, Schuler G (1999) An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods 223(1):77–92

    Article  CAS  PubMed  Google Scholar 

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Acknowledgement

We thank Maxine Swallow, Friederike Kruse, Melanie Gohmert, Martina Thiele, and Maike Hegemann for expert technical assistance and Dr. Katharina Lahl, Catharina Arnold-Schrauf, Christina Hesse, and Venkateswaran Ganesh for critical reading of the manuscript. C.T.M. was supported by the German National Academic Foundation. We would further like to thank the Cell Sorting Core Facility of the Hannover Medical School supported in part by the Braukmann-Wittenberg-Herz-Stiftung and Deutsche Forschungsgemeinschaft.

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Authors and Affiliations

  1. Institute of Infection Immunology, TWINCORE/Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), GmbH Feodor-Lynen-Str. 7, 30625, Hannover, Germany

    Christian Thomas Mayer & Tim Sparwasser

  2. The Rockefeller University, 1230 York Avenue, NY, 10065, New York, USA

    Christian Thomas Mayer

Authors
  1. Christian Thomas Mayer
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  2. Tim Sparwasser
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Corresponding author

Correspondence to Tim Sparwasser .

Editor information

Editors and Affiliations

  1. Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg University of Mainz, Mainz, Germany

    Ari Waisman

  2. University of Zurich Institute of Experimental Immunology, Zurich, Germany

    Burkhard Becher

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© 2014 Springer Science+Business Media New York

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Mayer, C.T., Sparwasser, T. (2014). Assessing the Suppressive Activity of Foxp3+ Regulatory T Cells. In: Waisman, A., Becher, B. (eds) T-Helper Cells. Methods in Molecular Biology, vol 1193. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1212-4_9

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  • DOI: https://doi.org/10.1007/978-1-4939-1212-4_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1211-7

  • Online ISBN: 978-1-4939-1212-4

  • eBook Packages: Springer Protocols

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